Transduction of dominant negative ATF-1 suppresses the pX gene expression in joint fibroblastic cells derived from HTLV-I transgenic rats

Ishizu, Akihiro; Tsuji, Takahiro; Abe, Asami; Saito, Saori; Takahashi, Toshiyuki; Ikeda, Hitoshi; Meruelo, Daniel; Yoshiki, Takashi

doi:10.1016/s0014-4800(02)00019-9

Cited by 6 publications

(10 citation statements)

References 28 publications

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“…For the vector control, lacZ/BB was similarly generated, using pSinRep/lacZ and DH-BB plasmids. We earlier confirmed that virus titers were comparable between ATF-1DN/BB and lacZ/BB [17]. To infect Sindbis virus vectors, cultured rat synovial cells (2 9 10 5 ) were exposed for 1 h at 37°C to a solution containing each vector (1 ml) and were then washed with PBS and incubated in the growth medium.…”

Section: Gene Transfection Into Cultured Rat Synovial Cellsmentioning

confidence: 59%

“…Primary culture of rat synovial cells Rat synovial cells were isolated from both nonarthritic joints of WKAH rats (n = 3) and affected joints of env-pX rats (n = 3), as previously described [17]. These cells were cultured in Roswell Park Memorial institute (RPMI) 1640 medium containing 10% fetal bovine serum (FBS) (Gibco BRL, Rockville, MD, USA).…”

Section: Immunohistochemistry For Crebmentioning

confidence: 99%

“…It is reported that high-titer proteins were generated from the genes transfected by Sindbis virus vector in various eukaryotic cells, including rat synovial cells [17][18][19]. The Sindbis virus vector carrying the ATF-1DN gene, ATF-1DN/BB, was constructed, as previously described [17]. Briefly, pSinRep5/ATF-1DN and a helper plasmid DH-BB (Invitrogen, Carlsbad, CA, USA), were linearized by XhoI.…”

Section: Gene Transfection Into Cultured Rat Synovial Cellsmentioning

confidence: 99%

“…For quantitative analysis of mRNA expression of IL-6, realtime reverse transcriptase polymerase chain reaction (RT-PCR) was performed using QuantiTect SYBR Green PCR (Qiagen, Hilden, Germany), as described [17]. Primers for IL-6 were 5 0 -attgaaaatctgctctggtcttctg-3 0 (sense) and 5 0 -ctggctttgtctttcttgttatcttg-3 0 (antisense).…”

Section: Gene Transfection Into Cultured Rat Synovial Cellsmentioning

confidence: 99%

“…These transgenic rats carrying the env-pX gene of human T-cell leukemia virus type-1 develop destructive arthritis with high titers of serum rheumatoid factor (RF) and are thus regarded as a suitable animal model of RA [9][10][11][12][13][14][15]. The dominant negative ATF-1 (ATF-1DN) contains multiple mutations in the DNA binding domain, though it conserves the leucine zipper motif [16,17]. ATF-1DN can bind to both CREB and ATF-1 and forms CREB/ATF-1DN and ATF-1/ATF-1DN heterodimers.…”

Section: Introductionmentioning

confidence: 99%

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Cyclic AMP response element-binding protein is implicated in IL-6 production from arthritic synovial cells

et al. 2009

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Overproduction of interleukin (IL)-6 from synovial cells is critically involved in the pathogenesis of rheumatoid arthritis (RA). Cyclic adenosine monophosphate (AMP) response element-binding protein (CREB), a leucine zipper transcription factor, is expressed at a high level in synovial cells of patients with RA. Although CREB transactivates IL-6 expression in vascular smooth muscle cells, the relation between CREB expression and IL-6 production from arthritic synovial cells remains unclear. In this study, to determine whether CREB is implicated in IL-6 production from arthritic synovial cells, a dominant negative molecule of activation transcription factor 1 (ATF-1) was transfected into synovial cells obtained from arthritic joints of env-pX rats. These transgenic rats carrying the env-pX gene of human T-cell leukemia virus type-1 develop destructive arthritis with high titers of serum rheumatoid factor and are thus regarded as a suitable model of RA. The dominant negative ATF-1 (ATF-1DN) constitutes a heterodimer with CREB and inhibits CREB function, as CREB/ATF-1DN heterodimers no longer bind to the target sequence of CREB. We showed that transfection of ATF-1DN significantly reduced IL-6 production from arthritic synovial cells. These findings suggest that CREB is implicated in IL-6 production from synovial cells and plays an important role in RA pathogenesis.

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Section: Gene Transfection Into Cultured Rat Synovial Cellsmentioning

confidence: 59%

Section: Immunohistochemistry For Crebmentioning

confidence: 99%

Section: Gene Transfection Into Cultured Rat Synovial Cellsmentioning

confidence: 99%

Section: Gene Transfection Into Cultured Rat Synovial Cellsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Cyclic AMP response element-binding protein is implicated in IL-6 production from arthritic synovial cells

et al. 2009

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Evidence of multiple infectious agents in mycosis fungoides lesions

Bonin

Tothova

Barbazza

et al. 2010

Experimental and Molecular Pathology

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Fluvastatin prevents the development of arthritis in env-pX rats via up-regulation of Rho GTPase-activating protein 12

Tanimura

Nishida

Horie

et al. 2020

Experimental and Molecular Pathology

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The pleiotropic effects of statins, including an antiarthritic potential, have been noted.This study aimed to determine the efficacy of statins on rheumatoid arthritis (RA) and clarify how statins affect its pathogenesis. Fluvastatin (500 µg/kg/day) or vehicle was given per os to env-pX rats, which carry the human T-cell leukemia virus type I env-pX gene and spontaneously develop destructive arthritis mimicking RA, for 30 days. Blood sampling and ultrasonography (US) of the ankle joints were conducted on days 0, 10, 20, and 30. On day 30, all rats were euthanized, and the ankle joints were subjected to histological analysis. To clarify how fluvastatin affects the pathogenesis of RA, comprehensive serum exosomal microRNA (miRNA) analysis was performed. Gene expression in the primary culture of synovial fibroblasts derived from arthritic rat and human and non-arthritic rat periarticular tissues was determined quantitatively by realtime reverse transcription-polymerase chain reaction (RT-PCR). As a result, the development of arthritis in env-pX rats was significantly suppressed by fluvastatin, which was evident from the viewpoints of serology, US imaging, and histology.Comprehensive serum exosomal miRNA analysis suggested that the expression of Rho GTPase-activating protein 12 (Arhgap12) was decreased in arthritic env-pX rats but increased with the administration of fluvastatin. Corresponding results were obtained by quantitative RT-PCR using primary culture of synovial fibroblasts. The collective findings suggest that fluvastatin prevents the development of arthritis in env-pX rats via the up-regulation of ARHGAP12. This study suggests that ARHGAP12 can be a possible therapeutic target of RA.

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Transduction of dominant negative ATF-1 suppresses the pX gene expression in joint fibroblastic cells derived from HTLV-I transgenic rats

Cited by 6 publications

References 28 publications

Cyclic AMP response element-binding protein is implicated in IL-6 production from arthritic synovial cells

Cyclic AMP response element-binding protein is implicated in IL-6 production from arthritic synovial cells

Evidence of multiple infectious agents in mycosis fungoides lesions

Fluvastatin prevents the development of arthritis in env-pX rats via up-regulation of Rho GTPase-activating protein 12

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